Ribonucleic acid (RNA), essential for protein production and immune function, undergoes glycosylation, a process that attaches glycans to RNA, generating unique glycoRNAs. These glycan-coated RNA molecules regulate immune responses and may be related to immune disorders. However, studying them is challenging due to RNA's fragility. Therefore, a robust method for identifying glycoRNA is important. To address this, we optimized parameters for RNA stability, oxidation, and digestion, thereby enriching and identifying glycoRNAs. This breakthrough paves the way for exploring their potential interactions with immune receptors and tumor suppression. Our approach involved investigating factors such as preservation reagent, enzyme buffer, digestion temperature, oxidant, glycosidase, and incubation time. We successfully optimized digestion conditions, achieving efficient cleavage of N-linked glycoRNAs at room temperature using 25 mM ammonium bicarbonate, demonstrating the effectiveness of this method. Additionally, RNA preservation in RNAlater at −80 °C allows controlled release of glycoRNAs within hours. While sequential digestion of different glycoRNA types is possible, significant degradation occurs after the first glycosidase step. Therefore, we recommend separate harvesting for each glycoRNA type. We also established RNA-seq analysis for identifying various glycoRNA types, including snoRNAs and tRNAs. The optimized SPCgRNA method paves the way for further research on N-glycosylation in health and disease.

Induction of podoplanin (PDPN) expression is a critical response of macrophages to LPS stimulation or bacterial infection in sepsis, but how this key process of TLR4-stimulated PDPN upregulation is regulated and the effect of PDPN expression on macrophage function remain elusive. Here, we determined how this process is regulated in vitro and in vivo. PDPN failed to be upregulated in TLR4-stimulated macrophages deficient in adhesion and degranulation-promoting adapter protein (ADAP), which could be rescued by the reconstitution of ADAP. A distinct PDPN ^hi peritoneal macrophage (PM) subset, which exhibited an M2-like phenotype and enhanced phagocytic activity, was generated in WT but not in ADAP-deficient septic mice. The blockade of PDPN ^hi PMs mimicked the effect of ADAP deficiency, which exacerbated sepsis. Mechanistically, Bruton’s tyrosine kinase–mediated (BTK-mediated) tyrosine phosphorylation of ADAP at Y ⁵⁷¹ worked together with mTOR to converge on STAT3 activation for the transactivation of the PDPN promoter. Moreover, agonist activation of STAT3 profoundly potentiated the PDPN ^hi PM subset generation and alleviated sepsis severity in mice. Together, our findings reveal a mechanism whereby ADAP resets macrophage function by controlling the TLR4-induced upregulation of PDPN as a host innate immune defense during sepsis.

Background: Colorectal cancer (CRC) ranks among the most prevalent malignant tumours of the digestive system globally and is associated with unfavourable survival outcomes. The exhaustion of CD8⁺ T cells serves a crucial role in facilitating tumour immune escape. Yet, the dynamic evolution of CD8⁺ T cell exhaustion and its impact on clinical prognosis across TNM (tumour‐node‐metastasis) stages in CRC remains incompletely characterized. Methods: Tumour and adjacent tissues (20 samples total) from 6 CRC patients spanning diverse TNM stages were analyzed using integrated single‐cell transcriptomic profiling (scRNA‐seq), single‐cell T cell receptor/B cell receptor sequencing (scVDJ‐seq), and spatial transcriptomics. T cell exhaustion markers, immune clonality, gene expression profiles, and the spatial distribution of both tumour cells and immune cells were systematically profiled. Functional enrichment and intercellular communication analyses were conducted. Key findings were validated using immunofluorescence and public datasets. Results: Our results illustrate how advancing TNM stages in CRC shape CD8⁺ T cell exhaustion through divergent TNFRSF18/CXCL13 dynamics and ribosomal stemness. TNFRSF18 expression was notably higher in T cells infiltrating tumour tissues relative to their counterparts in adjacent non‐tumorous areas, with high‐expressing CD8⁺ T cells exhibiting marked exhaustion features. During CRC progression, TNM‐stage‐driven remodelling of the tumour microenvironment (TME) induced progressive CD8⁺ T cell exhaustion marked by declining TNFRSF18 and rising CXCL13 expression in tumour‐infiltrating T cells elevation of both markers in the tumour compared with adjacent tissues. Moreover, we show that tumour cells displayed elevated expression of stemness‐associated ribosomal genes ( RPS7 , RPL8 , RPL30 ), peaking at stage T4, which correlated with poor prognosis and immune escape. Conclusions: This integrative multi‐omics study uncovers CD8⁺ T cell exhaustion dynamics and ribosomal stemness‐mediated immune evasion across CRC progression. CXCL13 , TNFRSF18 , and ribosomal proteins ( RPS7 / RPL8 / RPL30 ) are identified as novel biomarkers with direct prognostic value and therapeutic relevance, providing therapeutic targets for precision immunotherapy in CRC. Key points: Multi‐omics analysis reveals dynamic CD8 ⁺ T cell exhaustion patterns across CRC samples with different TNM stages. TNFRSF18 is highly expressed in exhausted tumour‐infiltrating CD8 ⁺ T cells and declines with disease progression. Ribosomal stemness in tumour cells promotes immune evasion by impairing TNF‐mediated CD8 ⁺ T cell function.

BACKGROUND Schizophrenia is a psychiatric disorder characterized by chronic or recurrent symptoms. Lurasidone was licensed in China in 2019 for the treatment of adult schizophrenia in adults with a maximum dose of 80 mg/d. However, post-market surveillance (PMS) with an adequate sample size is required for further validation of the drug’s safety profile and effectiveness. AIM To conduct PMS in real-world clinical settings and evaluate the safety and effectiveness of lurasidone in the Chinese population. METHODS A prospective, multicenter, open-label, 12-wk surveillance was conducted in mainland China. All patients with schizophrenia from 10 sites who had begun medication with lurasidone between September 2019 and August 2022 were eligible for enrollment. Safety assessments included adverse events (AEs), adverse drug reactions (ADRs), extrapyramidal symptoms (EPS), akathisia, use of EPS drugs, weight gain, and laboratory values as metabolic parameters and the QTc interval. The effectiveness was assessed using the brief psychiatric rating scale (BPRS) from baseline to the end of treatment. RESULTS A total of 965 patients were enrolled in the full analysis set and 894 in the safety set in this interim analysis. The average daily dose was 61.7 ± 19.08 mg (mean ± SD) during the treatment. AEs and ADRs were experienced by 101 patients (11.3%) and 78 patients (8.7%), respectively, which were mostly mild. EPS occurred in 25 individuals with a 2.8% incidence, including akathisia in 20 individuals (2.2%). Moreover, 59 patients received drugs for treating EPS during the treatment, with an incidence of 6.6% which dropped to 5.4% at the end of the treatment. The average weight change was 0.20 ± 2.36 kg (P = 0.01687) with 0.8% of patients showing a weight gain of ≥ 7% at week 12 compared with that at the baseline. The mean values of metabolic parameters and the QTc interval at baseline and week 12 were within normal ranges. The mean changes in total BPRS scores were -8.9 ± 9.76 (n = 959), -13.5 ± 12.29 (n = 959), and -16.8 ± 13.97 (n = 959) after 2/4, 6/8, and 12 wk, respectively (P < 0.001 for each visit compared with the baseline) using the last-observation-carried-forward method. CONCLUSION The interim analysis of the PMS of adult patients with schizophrenia demonstrate the safety and effectiveness of lurasidone in the Chinese population. No new safety or efficacy concerns were identified.

The protein kinesin family member 26B (KIF26B) is aberrantly expressed in various cancers. However, its particular role and association with tumor immune infiltration in colon adenocarcinoma (COAD) remain unclear. All original data were downloaded directly from The Cancer Genome Atlas (TCGA), UCSC Xena, and Gene Expression Omnibus (GEO) databases and processed with R 3.6.3. KIF26B expression was analyzed using Oncomine, TIMER, TCGA, GEO databases, and our clinical specimens. KIF26B expression at the protein level was explored with Human Protein Atlas (HPA) database. The upstream miRNAs and lncRNAs were predicted by StarBase and validated using RT-qPCR. Correlation of KIF26B expression with the expression of immune-related or immune checkpoint genes and GSEA analysis of KIF26B-related genes were investigated via R software. Relationship of KIF26B expression with immune biomarkers or tumor immune infiltration levels was studied through GEPIA2 and TIMER databases. KIF26B was upregulated, and its overexpression was closely related to overall survival (OS), disease-specific survival (DSS), progression-free interval (PFI), T stage, N stage, and CEA levels in COAD. MIR4435-2HG/hsa-miR−500a−3p/KIF26B axis was identified as the promising regulatory pathway of KIF26B. KIF26B expression was positively correlated with immune-related genes, tumor immune infiltration, and biomarker genes of immune cells in COAD, and KIF26B-related genes were significantly enriched in macrophage activation-related pathways. Expression of immune checkpoint genes, including PDCD1, CD274, and CTLA4, was also closely related to KIF26B expression. Our results clarified that ncRNA-based increased KIF26B expression was associated with a worse prognosis and high tumor immune infiltration in COAD.

Objective: To examine the relationship between the Dietary Inflammatory Index (DII), abnormal bowel habits, and systemic serum inflammatory markers.Methods: Data from 9,880 participants in the National Health and Nutrition Examination Survey (NHANES) 2005–2010 were analyzed. The DII was calculated from two 24-h dietary recalls. Bowel habits were assessed using the Bristol Stool Form Scale, and systemic inflammatory markers included AAPR, IBI, NLR, LMR, PNLR, LCR, LA, and PLR. Statistical analyses were performed using R, Zstats, and EmpowerStats to evaluate associations.Results: Higher DII scores were positively associated with abnormal bowel habits, including constipation [β (95% CI): 0.11 (0.01–0.22)] and diarrhea [β (95% CI): 0.42 (0.32–0.53)], and with PNLR [β (95% CI): 0.01 (0.01–0.01)], PNLRQ4 [β (95% CI): 0.13 (0.05–0.20)], IBI [β (95% CI): 0.02 (0.01–0.02)], and IBIQ4 [β (95% CI): 0.33 (0.25–0.42)] (p < 0.05). Negative associations were found with AAPR [β (95% CI): −0.33 (−0.60 - −0.06)] and AAPRQ4 [β (95% CI): −0.18 (−0.34 - −0.01)], while no significant associations were observed with LA, LCR, or LMR. Subgroup analyses confirmed stable associations between DII and both chronic diarrhea and constipation across seven subgroups. Smoothed curve fitting revealed nonlinear relationships. A J-shaped association between DII and chronic constipation was identified in BMI and IBI subgroups. For BMI >30, the breakpoint (K) was 1.89, with ORs of 1.228 (95% CI: 1.097–1.375) below and 3.318 (95% CI: 1.531–7.191) above this point. In the IBI Q4 subgroup, the breakpoint was 1.96, with ORs of 1.145 (95% CI: 1.013–1.294) below and 5.794 (95% CI: 2.359–14.228) above. In the diarrhea group, a U-shaped association was observed in the AAPR Q4 population, with a breakpoint of −1.312 and ORs of 0.657 (95% CI: 0.478–0.901) below and 1.266 (95% CI: 1.057–1.518) above.Conclusion: Higher DII scores are linked to an increased risk of chronic constipation and diarrhea and are associated with systemic inflammatory markers and factors such as BMI.